作　　者： ; ; ; ; ; ;

机构地区： 广西科学院

出　　处： 《生物技术通报》 2012年第7期170-175,共6页

摘　　要： 探究pflB、frdAB、fnr和AdhE四基因缺失突变株对大肠杆菌工程菌发酵生产异丁醇的影响。运用Red重组系统敲除大肠杆菌BW25113的pflB、frdAB、fnr和AdhE基因,构建pflB、frdAB、fnr和AdhE四基因缺失突变株E.coliBW25113H,结合本实验室已经构建的表达质粒pSTV29-alsS-ilvC-ilvD-kdcA,并检测该工程菌在1L发酵罐的发酵过程中的生物量、突变菌株的稳定性、异丁醇产量及有机酸含量的变化情况。成功获得pflB、frdAB、fnr和AdhE四基因缺失突变株BW25113H。发酵结果表明,该工程菌能以较长时间,较高比生长速率保持对数生长期,其稳定性较好,异丁醇产量增加了40%。成功构建pflB、frdAB、fnr和AdhE四基因缺失突变株BW25113H,结合非自身发酵途径使异丁醇的产量由3 g/L提升至4.2 g/L。 It was to study the effects of pflB, frdAB, fnr and AdhE four gene mutant disruptions on biosynthesis of isobutanol. The mutant E. coli BW25113H, which was deficient in pflB, frdAB, fnr and AdhE, was constructed by Red-combination technology, pSTV29-alsS-ilvC- ilvD-kdcA was introduced into engineered E. coli and investigate the biomass, stability, isobutanol production and the organic acid content in 1-L fermenter at 24 h. Compared with the parent strain E.coli BW25113, the mutant BW25113H was able to maintain higher growth rate at exponential phase, and the isobutanol production was increased by 40%. As the pflB, frdAB, fnr and AdhE four gene mutants BW25113H fermented with its non-fermentative pathway, the isobutanol production was enhanced from 3 g/L to 4.2 g/L. Furthermore, this work is associated with improvement of a non-natural host strain to facilitate production of isobutanol by engineering metabolic pathway.

关 键 词： 大肠杆菌工程菌 基因敲除 异丁醇 非发酵途径 重组系统

领　　域： [生物学]